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Temporal Gene Expression Profiles of Pre Blood-Fed Adult Females Immediately Following Eclosion in the Southern House Mosquito Culex Quinquefasciatus.

Reid WR, Zhang L, Liu N - Int. J. Biol. Sci. (2015)

Bottom Line: We found that adult females required a minimum of 48 h post-eclosion before they freely took their first blood meal.In addition, Trypsin was found to be up-regulated immediately following blood feeding, while trypsin and chymotrypsin were up-regulated at 48 h and 60 h post blood-feeding, respectively, suggesting that these proteases are likely involved in the digestion of the blood meal.Overall, this study reviewed multiple genes that might be involved in the adult female competency for blood meal acquisition in mosquitoes.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Entomology and Plant Pathology, Auburn University, Auburn, AL 36849, USA ; 2. Current address: UDSA-ARS Center for Medical Veterinary and Agricultural Entomology, Mosquito and Fly Research Unit, Gainesville, FL 32608, USA.

ABSTRACT
Prior to acquisition of the first host blood meal, the anautogenous mosquito Culex quinquefasciatus requires a period of time in order to prepare for the blood feeding and, later, vitellogenesis. In the current study, we conducted whole transcriptome analyses of adult female Culex mosquitoes to identify genes that may be necessary for both taking of the blood meal, and processing of the blood meal in adult female mosquitoes Cx. quinquefasciatus. We examined temporal expression of genes for the periods of post eclosion and prior to the female freely taking a blood meal. We further evaluated the temporal expression of certain genes for the periods after the taking of a blood meal to identify genes that may be necessary for both the taking of the blood meal, and the processing of the blood meal. We found that adult females required a minimum of 48 h post-eclosion before they freely took their first blood meal. We hypothesized that gene expression signatures were altered in the mosquitoes before blood feeding in preparation for the acquisition of the blood meal through changes in multiple gene expression. To identify the genes involved in the acquisition of blood feeding, we quantified the gene expression levels of adult female Cx. quinquefasciatus using RNA Seq throughout a pre-blooding period from 2 to 72 h post eclosion at 12 h intervals. A total of 325 genes were determined to be differentially-expressed throughout the pre-blooding period, with the majority of differentially-expressed genes occurring between the 2 h and 12 h post-eclosion time points. Among the up-regulated genes were salivary proteins, cytochrome P450s, odorant-binding proteins, and proteases, while the majority of the down-regulated genes were hypothetical or cuticular genes. In addition, Trypsin was found to be up-regulated immediately following blood feeding, while trypsin and chymotrypsin were up-regulated at 48 h and 60 h post blood-feeding, respectively, suggesting that these proteases are likely involved in the digestion of the blood meal. Overall, this study reviewed multiple genes that might be involved in the adult female competency for blood meal acquisition in mosquitoes.

No MeSH data available.


Distribution of differentially-expressed genes in adult sugar-fed female Culex quinquefasciatus for the initial 72h post-eclosion period.
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Figure 3: Distribution of differentially-expressed genes in adult sugar-fed female Culex quinquefasciatus for the initial 72h post-eclosion period.

Mentions: To further identify candidate genes that may be involved in preparing the female for the taking of a blood meal, we investigated the genes that were differentially-expressed throughout the time course investigated, i.e., 12, 24, 36, 48, 60, and 72 h time points after eclosion. Differential gene expression was determined as a time series, where gene differential expression was defined by comparing the expression levels of genes at any one time point with the expression at the prior time point; i.e., the expression at 24 h was compared with the expression at 12 h and so on. Overall, the majority of the genes that were identified as differentially up- and/or down-regulated occurred after 2 h post-eclosion and reached a maximum peak at 12 h, after which, a decrease in the number of differentially expressed genes was observed in each of the successive time points (Fig. 3, Table S2). A total of 325 genes were found to be differentially expressed, both up- and down-regulated genes, during 72 h post-eclosion. Roughly one-third of the differentially-expressed genes identified (101 genes) had no functional annotation in Vectorbase, while the remaining genes had predicted functions (Table S2). At 12 h post-eclosion, the greatest numbers of differentially-expressed genes was observed with 159 genes being up-regulated, and 74 down-regulated (Fig. 2). Among the up-regulated genes at 12 h post-eclosion were 32 salivary proteins and 2 apyrases, which may be involved in the prevention of platelet clotting during blood feeding 20, 10 cytochrome P450s (CPIJ005952, CPIJ011837, CPIJ010225, CPIJ010227, CPIJ000294, CPIJ019586, CPIJ019587, CPIJ020018, CPIJ012470, CPIJ010546 ), which were distributed among families CYP4, CYP6, CYP9, and CYP325, 20 proteases, and also genes involved in embryogenesis including wnt inhibitors and oskar 21, 22. Among the up-regulated genes at 24 h post-eclosion were 9 proteases and 1 olfactory receptor (CPIJ008023), which may be involved in preparation for the blood meal and host seeking, since both proteases and olfactory receptors have been linked to blood meal digestion and host seeking, respectively in mosquitoes 23-25. Among the genes down-regulated at 12 h post-eclosion were 27 hypothetical proteins, 11 cuticle proteins and 5 cytochrome P450 genes (CPIJ011841, CPIJ011840, CPIJ015954, CPIJ015961, and CPIJ015960), which were all in family CYP325. At 24 h post-eclosion, 32 hypothetical genes and 6 cuticle genes, which may be involved in the transition from the pharate to the adult, were down regulated.


Temporal Gene Expression Profiles of Pre Blood-Fed Adult Females Immediately Following Eclosion in the Southern House Mosquito Culex Quinquefasciatus.

Reid WR, Zhang L, Liu N - Int. J. Biol. Sci. (2015)

Distribution of differentially-expressed genes in adult sugar-fed female Culex quinquefasciatus for the initial 72h post-eclosion period.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4582154&req=5

Figure 3: Distribution of differentially-expressed genes in adult sugar-fed female Culex quinquefasciatus for the initial 72h post-eclosion period.
Mentions: To further identify candidate genes that may be involved in preparing the female for the taking of a blood meal, we investigated the genes that were differentially-expressed throughout the time course investigated, i.e., 12, 24, 36, 48, 60, and 72 h time points after eclosion. Differential gene expression was determined as a time series, where gene differential expression was defined by comparing the expression levels of genes at any one time point with the expression at the prior time point; i.e., the expression at 24 h was compared with the expression at 12 h and so on. Overall, the majority of the genes that were identified as differentially up- and/or down-regulated occurred after 2 h post-eclosion and reached a maximum peak at 12 h, after which, a decrease in the number of differentially expressed genes was observed in each of the successive time points (Fig. 3, Table S2). A total of 325 genes were found to be differentially expressed, both up- and down-regulated genes, during 72 h post-eclosion. Roughly one-third of the differentially-expressed genes identified (101 genes) had no functional annotation in Vectorbase, while the remaining genes had predicted functions (Table S2). At 12 h post-eclosion, the greatest numbers of differentially-expressed genes was observed with 159 genes being up-regulated, and 74 down-regulated (Fig. 2). Among the up-regulated genes at 12 h post-eclosion were 32 salivary proteins and 2 apyrases, which may be involved in the prevention of platelet clotting during blood feeding 20, 10 cytochrome P450s (CPIJ005952, CPIJ011837, CPIJ010225, CPIJ010227, CPIJ000294, CPIJ019586, CPIJ019587, CPIJ020018, CPIJ012470, CPIJ010546 ), which were distributed among families CYP4, CYP6, CYP9, and CYP325, 20 proteases, and also genes involved in embryogenesis including wnt inhibitors and oskar 21, 22. Among the up-regulated genes at 24 h post-eclosion were 9 proteases and 1 olfactory receptor (CPIJ008023), which may be involved in preparation for the blood meal and host seeking, since both proteases and olfactory receptors have been linked to blood meal digestion and host seeking, respectively in mosquitoes 23-25. Among the genes down-regulated at 12 h post-eclosion were 27 hypothetical proteins, 11 cuticle proteins and 5 cytochrome P450 genes (CPIJ011841, CPIJ011840, CPIJ015954, CPIJ015961, and CPIJ015960), which were all in family CYP325. At 24 h post-eclosion, 32 hypothetical genes and 6 cuticle genes, which may be involved in the transition from the pharate to the adult, were down regulated.

Bottom Line: We found that adult females required a minimum of 48 h post-eclosion before they freely took their first blood meal.In addition, Trypsin was found to be up-regulated immediately following blood feeding, while trypsin and chymotrypsin were up-regulated at 48 h and 60 h post blood-feeding, respectively, suggesting that these proteases are likely involved in the digestion of the blood meal.Overall, this study reviewed multiple genes that might be involved in the adult female competency for blood meal acquisition in mosquitoes.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Entomology and Plant Pathology, Auburn University, Auburn, AL 36849, USA ; 2. Current address: UDSA-ARS Center for Medical Veterinary and Agricultural Entomology, Mosquito and Fly Research Unit, Gainesville, FL 32608, USA.

ABSTRACT
Prior to acquisition of the first host blood meal, the anautogenous mosquito Culex quinquefasciatus requires a period of time in order to prepare for the blood feeding and, later, vitellogenesis. In the current study, we conducted whole transcriptome analyses of adult female Culex mosquitoes to identify genes that may be necessary for both taking of the blood meal, and processing of the blood meal in adult female mosquitoes Cx. quinquefasciatus. We examined temporal expression of genes for the periods of post eclosion and prior to the female freely taking a blood meal. We further evaluated the temporal expression of certain genes for the periods after the taking of a blood meal to identify genes that may be necessary for both the taking of the blood meal, and the processing of the blood meal. We found that adult females required a minimum of 48 h post-eclosion before they freely took their first blood meal. We hypothesized that gene expression signatures were altered in the mosquitoes before blood feeding in preparation for the acquisition of the blood meal through changes in multiple gene expression. To identify the genes involved in the acquisition of blood feeding, we quantified the gene expression levels of adult female Cx. quinquefasciatus using RNA Seq throughout a pre-blooding period from 2 to 72 h post eclosion at 12 h intervals. A total of 325 genes were determined to be differentially-expressed throughout the pre-blooding period, with the majority of differentially-expressed genes occurring between the 2 h and 12 h post-eclosion time points. Among the up-regulated genes were salivary proteins, cytochrome P450s, odorant-binding proteins, and proteases, while the majority of the down-regulated genes were hypothetical or cuticular genes. In addition, Trypsin was found to be up-regulated immediately following blood feeding, while trypsin and chymotrypsin were up-regulated at 48 h and 60 h post blood-feeding, respectively, suggesting that these proteases are likely involved in the digestion of the blood meal. Overall, this study reviewed multiple genes that might be involved in the adult female competency for blood meal acquisition in mosquitoes.

No MeSH data available.